RANCM: a new ranking scheme for assigning confidence levels to metabolite assignments in NMR-based metabolomics studies.

INTRODUCTION: The Metabolomics Standards Initiative has recommended four categories for metabolite assignments in NMR-based metabolic profiling studies. The "putatively annotated compound" category is most commonly reported by metabolomics investigators. However, there is significant ambiguity in reliability of "putatively annotated compound" assignments, which can range from low confidence made on minimal corroborating data to high confidence made on substantial corroborating data.
OBJECTIVES: To introduce a new ranking system, Rank and AssigN Confidence to Metabolites (RANCM), to assign confidence levels to "putatively annotated compound" assignments in NMR-based metabolic profiling studies.
METHODS: The ranking system was constructed with three confidence levels ranging from Rank 1 for the lowest confidence assignment level to Rank 3 for the highest confidence assignment level. A decision tree was constructed to guide rank selection for each metabolite assignment.
RESULTS: Examples are provided from experimental data demonstrating how to use the decision tree to make confidence level assignments to "putatively annotated compounds" in each of the three rank levels. A standard Excel sheet template is provided to facilitate decision-making, documentation and submission to data repositories.
CONCLUSION: RANCM is intended to reduce the ambiguity in "putatively annotated compound" assignments, to facilitate effective communication of the degree of confidence in "putatively annotated compound" assignments, and to make it easier for non-experts to evaluate the significance and reliability of NMR-based metabonomics studies. The system is straightforward to implement, based on the most common datasets collected in NMR-based metabolic profiling studies, and can be used with equal rigor and significance with any set of NMR datasets.